EP0596315B1 - Verfahren zum Binden einer Dialdehydstärke an eine Oberfläche, und nach dem Verfahren hergestellte Produkte - Google Patents

Verfahren zum Binden einer Dialdehydstärke an eine Oberfläche, und nach dem Verfahren hergestellte Produkte Download PDF

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Publication number
EP0596315B1
EP0596315B1 EP93116843A EP93116843A EP0596315B1 EP 0596315 B1 EP0596315 B1 EP 0596315B1 EP 93116843 A EP93116843 A EP 93116843A EP 93116843 A EP93116843 A EP 93116843A EP 0596315 B1 EP0596315 B1 EP 0596315B1
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Prior art keywords
gly
dialdehyde starch
das
solution
substrate
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French (fr)
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EP0596315A3 (en
EP0596315A2 (de
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Marie Dale Bryhan
Frances Marie Smith
Leroy S. Hersh
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Corning Inc
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Corning Inc
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
    • B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
    • B05D7/00—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials
    • B05D7/02—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials to macromolecular substances, e.g. rubber
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
    • C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
    • C03C17/28—Surface treatment of glass, not in the form of fibres or filaments, by coating with organic material
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
    • C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
    • C03C17/34—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions
    • C03C17/3405—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions with at least two coatings of organic materials
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
    • C08J7/04—Coating
    • C08J7/0427—Coating with only one layer of a composition containing a polymer binder
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D103/00—Coating compositions based on starch, amylose or amylopectin or on their derivatives or degradation products
    • C09D103/04—Starch derivatives
    • C09D103/10—Oxidised starch
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2403/00—Characterised by the use of starch, amylose or amylopectin or of their derivatives or degradation products

Definitions

  • This invention relates to methods of attaching dialdehyde starch to a surface and to products produced by those methods.
  • Cell culture products are used to grow large numbers of desired cells for medical testing, research and other uses.
  • Certain materials typically polymers, are optimal for use as substrates in cell culture products and are typically employed.
  • polystyrene is cheap, optically clear, and may be processed at low temperatures.
  • Large molecular weight cell attachment proteins (CAP's) are bound to substrate surfaces. Cells are then grown on the CAP-coated surfaces. The cells adhere to the CAP's, and the CAP's promote cell growth.
  • the CAP's bind readily to substrate materials such as polystyrene.
  • CAP's are very expensive and have short shelf lives, even at sub-freezing temperatures.
  • Dialdehyde starch is a commercially available compound made from starch by oxidation with periodic acid. DAS has been used in medicinal and bioactive compositions.
  • US Patent 3,495,000 discloses sustained release medicinal compositions adapted for peroral administration using a matrix comprising an admixture of dialdehyde starch and polyvinylpyrrolidone, polyvinyl chloride or ethyl cellulose. The medicament, dialdehyde starch, and polymer are mixed and granulated.
  • DAS contains two aldehyde groups per original glucose unit. These aldehyde groups readily react with molecules containing amine, imine, or hydrazide groups.
  • US Patent 3,706,633 discloses water-insoluble, enzymatically active compositions prepared by condensing dialdehyde starch with an alkylene diamine to produce a polymeric product, reducing and diazotizing the product, so as to produce a polymeric polydiazonium salt, and then coupling the product with an active enzyme.
  • US Patent 3,284,204 discloses an adhesive solution for bonding the hydrophobic film base and the hydrophilic emulsion in photographic film.
  • the adhesive solution comprises gelatin containing small amounts of an alkylene glycol derivative of polymannuronic acid and of a dialdehyde starch in water and a water-miscible solvent or mixture of solvents.
  • the alkylene glycol derivative of polymannuronic acid and the aldehyde starch are used to reduce so-called "after hardening" of the gelatin during storage and the specification makes it clear that the improvements which are sought in gelatin substratums are not achieved unless both the alkylene glycol derivative of polymannuronic acid and the aldehyde starch are used together.
  • the present invention satisfies the above described need.
  • the present invention provides a method of attaching dialdehyde starch to a polymer surface which does not contain chemical groups which will chemically react with dialdehyde starch, the method comprising: placing an aqueous solution of dialdehyde starch on the polymer surface for a period of time sufficient for at least a portion of the dialdehyde starch to be adsorbed by the surface; removing the solution; and drying the surface at a temperature of from 50°C to 150°C.
  • the present invention also provides a substrate to which substances may be coupled, the substrate comprising at least one polymer surface which does not contain chemical groups which will chemically react with dialdehyde starch, onto which polymer surface molecules of dialdehyde starch are adsorbed, thereby providing reactive aldehyde groups on the polymer surface available for coupling.
  • the present invention provides a substrate to which substances may be coupled obtainable by the method referred to above.
  • the present invention also provides products produced by coupling specific substances to the substrate of the present invention.
  • Biologically active molecules may be coupled to the substrate of the present invention without significantly deleteriously affecting their biological activity. These biologically active molecules include cell attachment proteins (CAP's) and cell attachment peptides (cap's). Coupling these molecules to the DAS-coated surfaces of the substrate of the present invention produces novel cell culture products.
  • CAP's cell attachment proteins
  • cap's cell attachment peptides
  • the products encompassed by the present invention are not limited to cell culture products, but encompasses any product produced by binding a substance or substances to the substrate of the present invention.
  • the present invention also provides a support for photographic film produced by coupling a gelatin film to the DAS-coated surfaces of a substrate of the present invention.
  • the method and products of the present invention can utilize a wide variety of polymers which are especially useful as cell culture substrate materials, including polystyrene, polypropylene, polyethylene terephthalate, polyallomer, cellulose acetate, and polymethylpentene.
  • the method and products of the present invention do not utilize surface materials containing hydroxyl groups or other chemical groups which will chemically react with DAS.
  • Figure 1 is a graph of the concentration of DAS solution used to coat an untreated polystyrene (UTPS) surface vs. the absorbance at 360 nanometers (nm) measured using the DNPH assay described herein on the coated surface. The absorbance is also translated into binding sites per nanometer squared (BS/nm 2 ).
  • Figure 2 is a graph of the boil time of the refluxing DAS solution vs. the absorbance at 240 nanometers of the solution. The graph also shows boil time vs. absorbance at 360 nm measured using the DNPH assay described herein on the coated surface.
  • Figure 3 is a graph of different batches of DAS solution boiled at a constant time vs. the absorbance at 240 nm of the solution. The graph also shows the batches vs. absorbance at 360 nm measured using the DNPH assay described herein on the coated surface.
  • Figure 4 is a graph of the time a polystyrene surface was soaked with 0.10 mg/ml DAS solution vs. the DNPH assay absorbance for the surface at 360 nm.
  • Figure 5 is a graph of the storage time in days at 30%, 70% and 90% relative humidity (RH) of a nonsterile DAS-coated polystyrene substrate produced with DAS using an 0.10 mg/ml DAS solution vs. the DNPH assay absorbance for the surface at 360 nm.
  • Figure 6 is a graph of the storage time in days at 30%, 70% and 90% relative humidity (RH) of a gamma radiation sterilized DAS-coated polystyrene substrate produced with DAS using an 0.10 mg/ml DAS solution vs. the DNPH assay absorbance for the surface at 360 nm.
  • Figure 7 is a graph of concentration of peptide suspension using the cell adhesion peptide Gly-Arg-Gly-Asp-Ser-Pro-Lys vs.
  • the Gold Assay absorbance at 530 nm of a peptide coated surface.
  • the surface is on a DAS-coated polystyrene substrate produced with a 0.10 mg/ml DAS solution.
  • the Gold Assay procedure is performed after peptide is adsorbed and loosely adsorbed peptide is washed away.
  • the Assay comprises adding colloidal gold solution to the coated surface and allowing the solution to stand for 2 to 4 hours at room temperature.
  • the spent gold solution is then removed, and the plate washed twice with distilled water.
  • the plate is then read at 530 nm in a microplate reader (DYNATECH MR5000).
  • Figures 8a, 8b and 8c are graphs of the growth time in fractions of days vs. the number of specific cells counted on the surface.
  • Figure 8a shows the results for rat aortic smooth muscle cells
  • Figure 8b shows the results for human tongue cells
  • Figure 8c shows the results for human endothelial cells.
  • a standard air-plasma or corona discharge treated polystyrene surface (Control)
  • a polystyrene surface coated with DAS using a 0.10 mg/ml solution and treated with a 1 mM Girard's reagent after binding the cell attachment peptide Gly-Arg-Gly-Asp-Ser-Pro-Lys (.1DAS/100R/G)
  • a polystyrene surface coated with DAS using a 0.10 mg/ml solution and treated with 4-hydrazinobenzene sulfonic acid after binding the cell attachment peptide Gly-Arg-Gly-Asp-Ser-Pro-Lys (0.1DAS/100R/H).
  • Figure 9 is a graph of growth time in days vs. human endothelial cells counted on the surface. Results for three surfaces are shown: a standard air-plasma or corona discharge treated polystyrene surface (Control), a standard air-plasma or corona discharge treated polystyrene surface with a fibronectin layer (Fibronectin), and a polystyrene surface coated with DAS using a 0.10 mg/ml solution and treated with a 1 mM Girard's reagent after binding the cell attachment peptide Gly-Arg Gly-Arg-Gly-Asp-Ser-Pro-Lys (.1DAS/100R/G).
  • Figure 10 is a graph of incubation at 20 days vs. the percent of wells in a culture plate showing no visible cell detachment. Results for three surfaces are shown: a standard air-plasma or corona discharge treated polystyrene surface (Control), a standard air-plasma or corona discharge treated polystyrene surface with a fibronectin layer (Fibronectin), and a polystyrene surface coated with DAS using a 0.10 mg/ml solution and treated with a 1 mM Girard's reagent after binding the cell attachment peptide Gly-Arg-Gly-Asp-Ser-Pro-Lys (.1DAS/100R/G).
  • Figure 11 is a graph of concentration of DAS solution used to coat a polystyrene substrate in mg/ml vs. antigenic activity of Immunoglobulin (IgG) chemically bound to a DAS-coated polystyrene (DAS/PS) substrate expressed as a percentage of the absorbance signal obtained from 1.5 micrograms per milliliter (ug/ml) IgG bound to a standard CORNING ELISA plate.
  • IgG Immunoglobulin
  • DAS/PS DAS-coated polystyrene
  • Results are shown for an untreated polystyrene sample bound to IgG using 0.02 M NaCNBH 3 [.02CN(O)], for an untreated polystyrene sample bound to IgG using 0.05 M NaCNBH 3 [.05CN(O)), and for a polystyrene sample treated with gamma radiation and bound to IgG using 0.05 M NaCNBH 3 [.05CN(E)]
  • an aqueous solution of DAS is produced by first making a paste.
  • the paste is made by adding a small amount of water to a weighed out portion of DAS.
  • the DAS paste is added to a suitable flask containing water for refluxing for a period of time sufficient to completely disperse the oxidized starch granules.
  • the clear DAS solution is then rapidly cooled to room temperature.
  • a certain volume of the DAS solution is then placed on the surface to be activated and allowed to stand for a period of time sufficient to allow the surface to adsorb at least a portion of DAS.
  • the DAS solution is then decanted from the vessel, and the ware is dried at 65°C to 70°C for a suitable period of time.
  • the DAS-coated surface produced by the method of the present invention may be coupled to a biologically-active molecule, such as a peptide or protein, by a variety of methods for surface attachment well known to those skilled-in-the-art.
  • a preferred method comprises placing a solution containing a reducing agent, a biologically active molecule and a suitable buffer at a pH of about 7 on the surface; gently shaking the solution-covered surface at a temperature of from 20°C to 25°C for at least 2 hours; removing the solution; and drying the surface at a temperature of from 35°C to 40°C for about 30 minutes.
  • the dissolution of DAS is a complex process involving dispersion, primary bond scission, and intramolecular reactions. Any method may be performed which provides an aqueous solution of DAS, including methods utilizing steam or microwaves. However, the above cited method utilizing refluxing is preferred.
  • the amount of DAS and water used may vary depending on the concentration of DAS desired. A DAS solution of 0.10 mg/ml of water (volume of 500 ml) requires about 30 minutes of boil time for an appearance of a clear solution.
  • the present invention is based on the discovery that DAS will strongly attach to various polymers and produce a reactive surface.
  • a two step process is required. First, the water dispersible DAS is physically adsorbed to the polymer surface. Then, after completely removing the DAS solution, the article is dried at moderate temperatures. A minimum amount of drying is necessary to produce an activated surface that is stable in an acid or neutral aqueous environment. DAS coated surfaces will degrade at pH values above 8.5.
  • the bonding of DAS to a cell growth surface as taught in the present invention occurs via physical bonds, possibly through a combination of hydrophobic and acid/base interactions. That is to say, the method of Ikada et al provides a hydroxyl containing surface to which DAS is chemically bound; the present invention provides a surface onto which molecules of DAS are adsorbed.
  • the present invention is applicable to a wide variety of surface materials, including polystyrene, polypropylene, polyethylene terephthalate, polyallomer, cellulose acetate, and polymethylpentene.
  • a key aspect of the present invention is that DAS becomes nearly irreversibly attached to some polymers after the moderate heating step.
  • a nonwetting surface such as polystyrene
  • the surface becomes wetting in seconds as DAS attaches.
  • DAS is easily rinsed off and the surface becomes nonwetting again unless the heating step is applied.
  • the surface is preferably dried at a temperature of from 65°C to 75°C.
  • the DAS used for the present invention have a high degree of oxidation.
  • DAS obtained from SIGMA (P9265) with a reported level of oxidation of 73% and containing 10% water.
  • Determination of the surface concentration of reactive aldehyde groups on DAS-coated surfaces was accomplished by using a modified version of a colorimetric method used for DAS solutions.
  • a chromophore, 2,4-dinitrophenylhydrazine (DNPH) (SIGMA D-2630) was dissolved in 1M HCl at a concentration of 0.25 mg/ml (0.9 mM) and then reacted with the DAS coated surface for 30 minutes in the dark.
  • the DNPH solution was then decanted, the surface washed with distilled water for 15 minutes and then dried in air.
  • the DNPH-DAS complex was solubilized by contact with 0.1 M NaOH for 5-10 minutes at room temperature in the dark. The pH was decreased to 2 after the NaOH soak and the absorbance was read at 360 nm.
  • Fig. 2 plots the DNPH absorbance values at 360 nm for DAS-coated surface vs. boil time of the DAS solution used to coat the surface. After clarity, the number of binding sites decrease with time, but at a much reduced rate of change. In order to obtain good reproducibility of binding sites of DAS coated polystyrene, the DAS solution boil is ended 5 minutes after clarity is obtained.
  • the low variability of the DNPH absorbance values at 360 nm for different batches of DAS coated polystyrene which were coated with DAS solutions boiled at a constant time is displayed in Fig. 3.
  • the A240 values (absorbance at 240 nm) of the DAS solution also showed good reproducibility even with the steady increase of A240 with boil time as shown in Fig. 2.
  • the rate of adsorption of DAS to polystyrene was determined by the DNPH assay of attached DAS. The results for an 0.10 mg/ml DAS solution are shown in Fig. 4. A minimum of about 5 minutes adsorption time is required for an equilibrium DNPH value. The minimum adsorption time increases to about 15 minutes for a concentration of 0.05 mg/ml DAS. The minimum adsorption time to reach an equilibrium DNPH value may vary depending on the surface used.
  • a large number of peptides may be bound to the DAS coated surfaces of the present invention. These include the peptides Gly-Arg-Gly-Asp-Ser-Pro-Lys, Lys-Gly, Gly-Gly-Tyr-Arg, and Arg-Lys-Asp-Val-Tyr.
  • a reducing agent preferably NaBH, or NaCNBH, was required to form a stable carbon to nitrogen bond.
  • Fig. 1 indicates a surface concentration of 3 BS per nm 2 at 0.10 mg/ml DAS solution concentration. If it is assumed that the peptide has similar reactivity as DNPH, a surface concentration of 1 microgram (ug) of peptide is calculated per well (96-well).
  • a preferred product of the present invention is an activated surface coupled to cell attachment peptides (cap's)
  • the present invention also encompasses an activated surface coupled to cell attachment proteins (CAP's).
  • IgG immunoglobulin
  • An application would be a coating for the 96-well plate used as a support for enzyme immunoassays (ELISA).
  • Fig. 11 is a plot of data obtained from the determination of the antigenic activity of IgG chemically bound to DAS-coated polystyrene.
  • Results are shown for an untreated polystyrene sample bound to IgG using 0.02 M NaCNBH 3 [.02CN(O)], for an untreated polystyrene sample bound to IgG using 0.05 M NaCNBH 3 [.05CN(O)], and for a polystyrene sample treated with gamma radiation and bound to IgG using 0.05 M NaCNBH 3 [.05CN(E)].
  • the results are expressed as a percentage of the signal obtained with IgG physically bound to a standard CORNING ELISA plate.
  • the maximum response of the DAS-coated polystyrene was just about equal to the control surface - a gamma radiation treated polystyrene ELISA surface.
  • Bovine serum albumin and bovine gelatin have also been successfully bound to DAS-coated polystyrene surfaces of the present invention.
  • Figs. 8a-10 display results of growth studies performed at the National Institute of Health laboratories with various primary cell lines grown on the Gly-Arg-Gly-Asp-Ser-Pro-Lys/DAS/polystyrene surface. The results in these cases show that after 3 to 6 days, the surface with bound Gly-Arg-Gly-Asp-Ser-Pro-Lys peptide had nearly two times the growth obtained with the control surface.
  • the control surface was a standard air-plasma or corona treated polystyrene tissue culture surface.
  • Fig. 9 shows some growth studies performed with primary human endothelial cells (HEC).
  • HEC primary human endothelial cells
  • the peptide/DAS surface showed about 50% more cells than the standard air-plasma or corona treated polystyrene tissue culture surface used as control. More importantly, this plot indicates that the peptide-coated surface of the present invention promoted about 15% more cell growth than a standard air-plasma or corona treated polystyrene tissue culture surface having a fibronectin layer - the present state-of-the-art.
  • the following table 2 lists the performance of various cells grown on a Gly-Arg-Gly-Asp-Ser-Pro-Lys/DAS/polystyrene surface of the present invention as a percentage of growth seen on a control surface - a standard air-plasma or corona treated polystyrene tissue culture surface.
  • peptide surface in prolonging the viability of a bound confluent monolayer of the human endothelial cells (HEC's) as shown in Fig. 10. There was a 40% increase in the number of wells showing no visible cell detachment after a 20 day incubation in cell growth media when comparing the peptide surface with a fibronectin coated surface.
  • a paste was made by adding about 0.050 g DAS to about 2 ml of water in a plastic weighing boat.
  • the DAS paste was added to a 1000 ml boiling flask and water was added to a total volume of about 500 ml.
  • the boiling flask was fitted with a reflux condenser and heated for about 53 minutes on a hot plate turned to the high setting.
  • the solution cleared after about 25 minutes of boiling (about 48 minutes total heating time).
  • the flask was then cooled in an ice-bath for about 15 minutes.
  • a solution of 0.10 mg/ml IgG in 100 mM MOPS and 20 mM NaCNBH 3 at about pH 7.5 and 100 mM acetate at about pH 4.5 was produced. 2 ml of the solution was added to each well of a 24 well plate. The plate was placed on a rocking shaker table and allowed to stand for about 18 hours at about 22°C. The solution was then removed by pipette. The wells were then washed with phosphate-buffered saline at about pH 7. The plate was then dried at about 30°C to 35°C for about 1 hour in a forced air oven (Fisher Isotemp).
  • the liquid and unattached cells were removed from the wells with pipettes. 2 ml of phosphate buffered saline (PBS) was then added to each well and the wells were shaken gently. The liquid was then removed from the wells. The PBS wash step was then repeated. 1 ml of 0.1% trypsin was then added to each well. After 10 minutes, the released cells were counted in a Coulter counter.
  • PBS phosphate buffered saline

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Claims (11)

  1. Verfahren zum Anbringen von Dialdehydstärke an eine Polymeroberfläche, die keine chemischen Gruppen enthält, die chemisch mit Dialdehydstärke reagieren, wobei das Verfahren die nachfolgenden Schritte umfaßt:
    Aufbringen einer wässerigen Dialdehydstärkelösung auf die Polymeroberfläche für eine Zeitdauer, die ausreichend ist, daß zumindest ein Teil der Dialdehydstärke von der Oberfläche adsorbiert wird;
    Entfernen der Lösung; und
    Trocknen der Oberfläche bei einer Temperatur von 50°C-150°C.
  2. Verfahren nach Anspruch 1, wobei die wässerige Dialdehydstärkelösung durch ein Verfahren mit den nachfolgenden Schritten hergestellt wird:
    Zugabe von Wasser zu einem Teil der Dialdehydstärke, um eine Paste auszubilden;
    Refluxieren der Paste mit Wasser, bis die oxidierten Stärkekörnchen ausreichend im Wasser dispergiert sind; und
    schnelles Abkühlen der Dialdehydstärkelösung auf Raumtemperatur.
  3. Verfahren nach Anspruch 1 oder 2, wobei die wässerige Dialdehydstärkelösung eine Konzentration von zumindest 0,05 mg/ml aufweist und die Polymeroberfläche der wässerigen Dialdehydstärkelösung zumindest 15 Minuten lang ausgesetzt wird, oder eine Konzentration von zumindest 0,10 mg/ml aufweist und die Polymeroberfläche der wässerigen Dialdehydstärkelösung zumindest fünf Minunten lang ausgesetzt wird, und/oder wobei die Oberfläche bei einer Temperatur von 65°C-75°C getrocknet wird.
  4. Verfahren nach einem der Ansprüche 1-3, wobei die Polymeroberfläche aus Polystyrol, Polypropylen, Polyethylenterephthalat, Polyallomer, Celluloseacetat und Polymethylpenten ausgewählt wird, wobei die Oberfläche wahlweise die Innenseite eines Laborbehälters darstellt.
  5. Verfahren zur Herstellung einer Oberfläche für wachsende Zellen, wobei das Verfahren die nachfolgenden Schritte umfaßt:
    Aufbringen einer Lösung, die ein Reduktionsmittel, biologisch aktive Moleküle und einen geeigneten Puffer mit einem pH-Wert von etwa 7 enthält, auf eine Oberfläche, die durch das Verfahren nach einem der Ansprüche 1-4 aktiviert wurde;
    sanftes Schütteln der mit der Lösung bedeckten Oberfläche bei einer Temperatur von 20°C-25°C für zumindest zwei Stunden;
    Entfernen der Lösung und
    Trocknen der Oberfläche bei einer Temperatur von 35°C-40°C.
  6. Verfahren nach Anspruch 5, wobei die biologisch aktiven Moleküle aus Gly-Arg-Gly-Asp-Ser-Pro-Lys, Lys-Gly, Gly-Gly-Tyr-Arg und Arg-Lys-Asp-Val-Tyr ausgewählt werden.
  7. Substrat, an die Substanzen angekoppelt werden können, wobei das Substrat zumindest eine Polymeroberfläche aufweist, die keine chemischen Gruppen enthält, die chemisch mit Dialdehydstärke reagieren, wobei auf die Polymeroberfläche Moleküle aus Dialdehydstärke adsorbiert werden, wodurch reaktive Aldehydgruppen zum Koppeln auf der Polymeroberfläche zur Verfügung gestellt werden.
  8. Substrat mit biologisch aktiven Molekülen, die an seine Oberfläche gekoppelt sind, umfassend ein Substrat nach Anspruch 7 mit biologisch aktiven Molekülen, die an die reaktiven Aldehydgruppen der mit Dialdehydstärke beschichteten Oberfläche bzw. den mit Dialdehydstärke beschichteten Oberflächen gekoppelt sind.
  9. Substrat nach Anspruch 8, wobei die biologisch aktiven Moleküle Zellbindungspeptide oder Zellbindungsproteine sind.
  10. Substrat nach Anspruch 9, wobei die Zellbindungspeptide aus Gly-Arg-Gly-Asp-Ser-Pro-Lys, Lys-Gly, Gly-Gly-Tyr-Arg und Arg-Lys-Asp-Val-Tyr ausgewählt sind oder die Zellbindungsproteine aus Immunglobulinen und Rinderserumalbumin ausgewählt werden.
  11. Mit Gelatine beschichtetes Substrat, umfassend ein Substrat nach Anspruch 7 mit einem Gelatinefilm, der an Dialdehydgruppen der mit Dialdehydstärke beschichteten Oberfläche (n) gekoppelt ist.
EP93116843A 1992-11-05 1993-10-19 Verfahren zum Binden einer Dialdehydstärke an eine Oberfläche, und nach dem Verfahren hergestellte Produkte Expired - Lifetime EP0596315B1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US07/972,327 US5281660A (en) 1992-11-05 1992-11-05 Method of attaching dialdehyde starch to a surface and products produced by that method
US972327 1992-11-05

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EP0596315A2 EP0596315A2 (de) 1994-05-11
EP0596315A3 EP0596315A3 (en) 1994-07-20
EP0596315B1 true EP0596315B1 (de) 1998-01-28

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EP (1) EP0596315B1 (de)
JP (1) JPH06209759A (de)
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DE (1) DE69316707T2 (de)

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US5931165A (en) * 1994-09-06 1999-08-03 Fusion Medical Technologies, Inc. Films having improved characteristics and methods for their preparation and use
US6391655B1 (en) * 1997-07-30 2002-05-21 Corning Incorporated Oxidized styrenic polymers for DNA binding
US6615842B1 (en) 1998-02-13 2003-09-09 Cerami Consulting Corp. Methods for removing nucleophilic toxins from tobacco smoke
AU759687B2 (en) * 1998-07-21 2003-04-17 Pharmexa A/S Coating of solid surfaces with activated polyhydroxypolymers
US7041506B2 (en) * 2001-11-19 2006-05-09 Becton Dickinson And Company Peptides promoting cell adherence, growth and secretion
US7384742B2 (en) * 2002-08-16 2008-06-10 Decision Biomarkers, Inc. Substrates for isolating reacting and microscopically analyzing materials
WO2009111022A1 (en) * 2008-03-07 2009-09-11 Corning Incorporated Hydrophobically modified cellulose ether derivatives for cell culture and release

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EP0166998A2 (de) * 1984-06-04 1986-01-08 TERUMO KABUSHIKI KAISHA trading as TERUMO CORPORATION Medizinisches Werkzeug und Verfahren zur Herstellung

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Also Published As

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US5563215A (en) 1996-10-08
DE69316707D1 (de) 1998-03-05
EP0596315A3 (en) 1994-07-20
CA2108608A1 (en) 1994-05-06
JPH06209759A (ja) 1994-08-02
EP0596315A2 (de) 1994-05-11
US5281660A (en) 1994-01-25
DE69316707T2 (de) 1998-05-14

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